Fig. 9

Binding of WRKY50BD to the sequences S1 and S2 requires the WT-boxes CGACTTTT. Lanes 1: free probe. Lanes 2: free probe plus purified protein extract of E. coli not expressing WRKY50BD. Lanes 3: free probe plus purified WRKY50BD. Lanes 4: free probe, purified WRKY50BD, and unlabelled competitor (S1 or S2, respectively) in a 4000-fold molar excess. Lanes 5: free probe, purified WRKY50BD, and unlabelled mutations S1mut1 or S2mut1, respectively in a 4000-fold molar excess. The sequences of S1, S1mut1, S2, and S2mut1 are shown. Unchanged nucleotides in S1mut1 and S2mut1 are indicated by a dash (−). A P designates the position of the free probe and a specific DNA-protein complex is marked with an asterisk (*). The gel shifts were done three times and one example is shown. The original unprocessed figures are shown in Fig. S1 and Fig. S2